RNAseq of E2A-PBX1⁺/preBCR⁺ leukemia CNS-infiltrating cells compared across treatments. Bulk RNA sequencing of FACS-sorted CNS-infiltration E2A-PBX1-GFP⁺ leukemic cells from vehicle- (n = 3), dasatinib- (n = 3), ibrutinib- (n = 3), and dasatinib + ibrutinib–treated mice (n = 3) was performed. (A) MA plot displaying 349 most significant genes (P < .05) between all the treatments. (B) Heat map showing BCR–associated differential expressed (DE) genes. Myc (P = .06) is downregulated in the double-treated mice compared with vehicle-treated mice. A trend for decreased expression of BCR-associated genes Lck, Zap70, and Cd79a showed a trend for decreased expression under the treatment with BTKi alone or in combination with dasatinib. In contrast, Pten (P < .05) and Mapk1/ERK2 (P < .05) are upregulated in the double-treated mice compared with vehicle-treated mice. (C) Venn diagrams showing the most significant in-common up- and downregulated genes between the treatments. (D) Validation of Pten, Myc, and Mapk1 genes by RT-qPCR. Actb was used as a housekeeping gene. The experiment was performed in triplicate. Bars represent the mean and error bars, the SEM, of 3 independent experiments.